CA2738635C - Continuous directed evolution of proteins and nucleic acids - Google Patents

Continuous directed evolution of proteins and nucleic acids Download PDF

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Publication number
CA2738635C
CA2738635C CA2738635A CA2738635A CA2738635C CA 2738635 C CA2738635 C CA 2738635C CA 2738635 A CA2738635 A CA 2738635A CA 2738635 A CA2738635 A CA 2738635A CA 2738635 C CA2738635 C CA 2738635C
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Prior art keywords
cas
phage
gene
phagemid
host cells
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Expired - Fee Related
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CA2738635A
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English (en)
French (fr)
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CA2738635A1 (en
Inventor
David R. Liu
Kevin M. Esvelt
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Harvard College
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Harvard College
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Priority to CA 3059768 priority Critical patent/CA3059768A1/en
Priority to CA3073384A priority patent/CA3073384A1/en
Publication of CA2738635A1 publication Critical patent/CA2738635A1/en
Application granted granted Critical
Publication of CA2738635C publication Critical patent/CA2738635C/en
Expired - Fee Related legal-status Critical Current
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1058—Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102—Mutagenizing nucleic acids
    • C12N15/1024—In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1037—Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Plant Pathology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Ecology (AREA)
  • Virology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
CA2738635A 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids Expired - Fee Related CA2738635C (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CA 3059768 CA3059768A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids
CA3073384A CA3073384A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US9466608P 2008-09-05 2008-09-05
US61/094,666 2008-09-05
PCT/US2009/056194 WO2010028347A2 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids

Related Child Applications (2)

Application Number Title Priority Date Filing Date
CA 3059768 Division CA3059768A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids
CA3073384A Division CA3073384A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids

Publications (2)

Publication Number Publication Date
CA2738635A1 CA2738635A1 (en) 2010-03-11
CA2738635C true CA2738635C (en) 2019-12-31

Family

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Family Applications (3)

Application Number Title Priority Date Filing Date
CA3073384A Abandoned CA3073384A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids
CA2738635A Expired - Fee Related CA2738635C (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids
CA 3059768 Pending CA3059768A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CA3073384A Abandoned CA3073384A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids

Family Applications After (1)

Application Number Title Priority Date Filing Date
CA 3059768 Pending CA3059768A1 (en) 2008-09-05 2009-09-08 Continuous directed evolution of proteins and nucleic acids

Country Status (5)

Country Link
US (2) US9023594B2 (ja)
EP (2) EP2342336B1 (ja)
JP (3) JP5723774B2 (ja)
CA (3) CA3073384A1 (ja)
WO (1) WO2010028347A2 (ja)

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WO2017015559A2 (en) 2015-07-23 2017-01-26 President And Fellows Of Harvard College Evolution of bt toxins
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Also Published As

Publication number Publication date
JP2016104033A (ja) 2016-06-09
WO2010028347A3 (en) 2010-08-19
JP2014237724A (ja) 2014-12-18
EP2342336A4 (en) 2012-05-02
CA2738635A1 (en) 2010-03-11
CA3073384A1 (en) 2010-03-11
JP5723774B2 (ja) 2015-05-27
EP2342336A2 (en) 2011-07-13
JP6050295B2 (ja) 2016-12-21
JP2012501656A (ja) 2012-01-26
US9771574B2 (en) 2017-09-26
WO2010028347A2 (en) 2010-03-11
EP3199630B1 (en) 2019-05-08
EP3199630A1 (en) 2017-08-02
CA3059768A1 (en) 2010-03-11
US20150275202A1 (en) 2015-10-01
US20110177495A1 (en) 2011-07-21
EP2342336B1 (en) 2016-12-14
US9023594B2 (en) 2015-05-05

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